LOCUS       sequence-474681-        7082 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express BCL6-LgBiT Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7082)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: BCL6-LgBiT Vector
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2823..2904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2860..2879)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2976..3333
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2976..3167
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2999..3019)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3184..3319
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3364..4401
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4380..4388
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4425..4473
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4623..4640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4757..5302)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4837..4856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5519..6313)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5630..5649)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6240..6259
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6345..6392)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(6425..6445)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6600..6903
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7023..7041
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7023..7040
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gcctcgccgg ctgacagctg tatccagttc acccgccatg ccagtgatgt tcttctcaac
       61 cttaatcgtc tccggagtcg agacatcttg actgatgttg tcattgttgt gagccgtgag
      121 cagtttagag cccataaaac ggtcctcatg gcctgcagtg gcctgttcta tagcatcttt
      181 acagaccagt tgaaatgcaa ccttagtgtg atcaatctag atcctgagat caaccctgag
      241 ggattctgca tcctcctgga cttcatgtac acatctcggc tcaatttgcg ggagggcaac
      301 atcatggctg tgatggccac ggctatgtac ctgcagatgg agcatgttgt ggacacttgc
      361 cggaagttta ttaaggccag tgaagcagag atggtttctg ccatcaagcc tcctcgtgaa
      421 gagttcctca acagccggat gctgatgccc caagacatca tggcctatcg gggtcgtgag
      481 gtggtggaga acaacctgcc actgaggagc gcccctgggt gtgagagcag agcctttgcc
      541 cccagcctgt acagtggcct gtccacaccg ccagcctctt attccatgta cagccacctc
      601 cctgtcagca gcctcctctt ctccgatgag gagtttcggg atgtccggat gcctgtggcc
      661 aaccccttcc ccaaggagcg ggcactccca tgtgatagtg ccaggccagt ccctggtgag
      721 tacagccggc cgactttgga ggtgtccccc aatgtgtgcc acagcaatat ctattcaccc
      781 aaggaaacaa tcccagaaga ggcacgaagt gatatgcact acagtgtggc tgagggcctc
      841 aaacctgctg ccccctcagc ccgaaatgcc ccctacttcc cttgtgacaa ggccagcaaa
      901 gaagaagaga gaccctcctc ggaagatgag attgccctgc atttcgagcc ccccaatgca
      961 cccctgaacc ggaagggtct ggttagtcca cagagccccc agaaatctga ctgccagccc
     1021 aactcgccca cagagtcctg cagcagtaag aatgcctgca tcctccaggc ttctggctcc
     1081 cctccagcca agagccccac tgaccccaaa gcctgcaact ggaagaaata caagttcatc
     1141 gtgctcaaca gcctcaacca gaatgccaaa ccagaggggc ctgagcaggc tgagctgggc
     1201 cgcctttccc cacgagccta cacggcccca cctgcctgcc agccacccat ggagcctgag
     1261 aaccttgacc tccagtcccc aaccaagctg agtgccagcg gggaggactc caccatccca
     1321 caagccagcc ggctcaataa catcgttaac aggtccatga cgggctctcc ccgcagcagc
     1381 agcgagagcc actcaccact ctacatgcac cccccgaagt gcacgtcctg cggctctcag
     1441 tccccacagc atgcagagat gtgcctccac accgctggcc ccacgttccc tgaggagatg
     1501 ggagagaccc agtctgagta ctcagattct agctgtgaga acggggcctt cttctgcaat
     1561 gagtgtgact gccgcttctc tgaggaggcc tcactcaaga ggcacacgct gcagacccac
     1621 agtgacaaac cctacaagtg tgaccgctgc caggcctcct tccgctacaa gggcaacctc
     1681 gccagccaca agaccgtcca taccggtgag aaaccctatc gttgcaacat ctgtggggcc
     1741 cagttcaacc ggccagccaa cctgaaaacc cacactcgaa ttcactctgg agagaagccc
     1801 tacaaatgcg aaacctgcgg agccagattt gtacaggtgg cccacctccg tgcccatgtg
     1861 cttatccaca ctggtgagaa gccctatccc tgtgaaatct gtggcacccg tttccggcac
     1921 cttcagactc tgaagagcca cctgcgaatc cacacaggag agaaacctta ccattgtgag
     1981 aagtgtaacc tgcatttccg tcacaaaagc cagctgcgac ttcacttgcg ccagaagcat
     2041 ggcgccatca ccaacaccaa ggtgcaatac cgcgtgtcag ccactgacct gcctccggag
     2101 ctccccaaag cctgcgtttc tcaaggcagt tcaggtggtg gcgggagcgg aggtggaggc
     2161 tcgagcggtg tcttcacact cgaagatttc gttggggact gggaacagac agccgcctac
     2221 aacctggacc aagtccttga acagggaggt gtgtccagtt tgctgcagaa tctcgccgtg
     2281 tccgtaactc cgatccaaag gattgtccgg agcggtgaaa atgccctgaa gatcgacatc
     2341 catgtcatca tcccgtatga aggtctgagc gccgaccaaa tggcccagat cgaagaggtg
     2401 tttaaggtgg tgtaccctgt ggatgatcat cactttaagg tgatcctgcc ctatggcaca
     2461 ctggtaatcg acggggttac gccgaacatg ctgaactatt tcggacggcc gtatgaaggc
     2521 atcgccgtgt tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa
     2581 attatcgacg agcgcctgat cacccccgac ggctccatgc tgttccgagt aaccatcaac
     2641 agctaaggcc gcgactctag aaacacctag gctctcgtac gaagtcagct gataacgtcg
     2701 acgtattgcg gccgcggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2761 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2821 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2881 caaataaagc atttttttca ctgccgacgt cacgcgtctt gcgcagcacc atggcctgaa
     2941 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     3001 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3061 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3181 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3241 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3301 ggaggctttt ttggaggcct aggcttttgc aaaaagctcg attcttctga cactagcgcc
     3361 accatgaaga agcccgaact caccgctacc agcgttgaaa aatttctcat cgagaagttc
     3421 gacagtgtga gcgacctgat gcagttgtcg gagggcgaag agagccgagc cttcagcttc
     3481 gatgtcggcg gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg cttctacaaa
     3541 gaccgctacg tgtaccgcca cttcgccagc gctgcactac ccatccccga agtgttggac
     3601 atcggcgagt tcagcgagag cctgacatac tgcatcagta gacgcgccca aggcgttact
     3661 ctccaagacc tccccgaaac agagctgcct gctgtgttac agcctgtcgc cgaagctatg
     3721 gatgctattg ccgccgccga cctcagtcaa accagcggct tcggcccatt cgggccccaa
     3781 ggcatcggcc agtacacaac ctggcgggat ttcatttgcg ccattgctga tccccatgtc
     3841 taccactggc agaccgtgat ggacgacacc gtgtccgcca gcgtagctca agccctggac
     3901 gaactgatgc tgtgggccga agactgtccc gaggtgcgcc acctcgtcca tgccgacttc
     3961 ggcagcaaca acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga ctggtccgaa
     4021 gctatgttcg gggacagtca gtacgaggtg gccaacatct tcttctggcg gccctggctg
     4081 gcttgcatgg agcagcagac tcgctacttc gagcgccggc atcccgagct ggccggcagc
     4141 cctcgtctgc gagcctacat gctgcgcatc ggcctggatc agctctacca gagcctcgtg
     4201 gacggcaact tcgacgatgc tgcctgggct caaggccgct gcgatgccat cgtccgcagc
     4261 ggggccggca ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt atggaccgac
     4321 ggctgcgtcg aggtgctggc cgacagcggc aaccgccggc ccagtacacg accgcgcgct
     4381 aaggaggtag gtcgagttta gactctagcc atcacgatgg ccgcaataaa atatctttat
     4441 tttcattaca tctgtgtgtt ggttttttgt gtgacgattc cggacatcag cgctagcgga
     4501 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     4561 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     4621 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     4681 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     4741 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     4801 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4861 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     4921 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     4981 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     5041 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     5101 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     5161 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     5221 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     5281 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat
     5341 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     5401 gtaattctca tgtttgacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5461 attaccgcct ttgagtgagc tgataccggt tgggcgtcgc ttggtcggtc atttcgaatc
     5521 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     5581 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     5641 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     5701 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     5761 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     5821 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     5881 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     5941 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     6001 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     6061 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     6121 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     6181 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     6241 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     6301 cttgttcaat catgatctgc ctcctgggtt tagttcctca ccttgtcgta ttatactatg
     6361 ccgatatact atgccgatga ttaattgtca acagtattag aattcgattt gacctcccac
     6421 cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca ttttggaaag
     6481 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     6541 aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc
     6601 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc
     6661 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga
     6721 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     6781 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6841 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6901 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     6961 ctagtggcgc gccagggagg cctatataag cagagctggt ttagtgaacc gtcagatcgc
     7021 ccatttaggt gacactatag aatacaagct acttgttctt tttgcagcta gcgatcgcca
     7081 tg
//
